期刊
BIOSENSORS & BIOELECTRONICS
卷 36, 期 1, 页码 129-134出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2012.04.013
关键词
Interferon-gamma; Aptamer; Electrochemical sensor; Hybridization chain reaction
类别
资金
- NSFC [21025521, 21035001, 21190041]
- National Key Basic Research Program [2011CB911000]
- CSIRT Program
- NSF of Hunan Province [10JJ7002]
- Scientific Research Fund of Hunan Provincial Education Department [08A065]
A novel electrochemical aptasensor based on hybridization chain reaction (HCR) with enzyme-signal amplification was constructed for the detection of interferon-gamma (IFN-gamma). In this aptasensor, the recognition probes which contained the sequence of IFN-gamma aptamer were initially binded to IFN-gamma. and the unbound recognition probes were captured on the electrode as an initiator to trigger the HCR. The two DNA hairpins bio-H1 and bio-H2 were opened by the recognition probe, and bound one by one on the electrode. The biotin was used as a tracer in the hairpins and streptavidin-alkaline phosphatase (SA-ALP) as a reporter molecule. Then, SA-ALP converted its electro-inactive substrate 1-naphthyl phosphate into an electroactive derivative 1-naphthol generating amplified electrochemical signal by differential pulse voltammetry (DPV). The activity of the immobilized enzyme was voltammetrically determined by measuring the amount of 1-naphthol generated for enzymatic dephosphorylation of 1-naphthyl phosphate. The electrochemical signal observed was inversely related to the concentration of IFN-gamma. The proposed approach showed a high sensitivity for IFN-gamma in a concentration range of 0.5-300 nM with a detection limit of 0.3 nM. The sensing system also provided satisfactory results for the detection of IFN-gamma in the cell media. (C) 2012 Elsevier B.V. All rights reserved.
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